Obesity-linked (type-2) diabetes exacts a significant health toll, but there are few disease-modifying treatments to stem this "diabesity epidemic." Better understanding of the mechanisms that regulate energy homeostasis are needed to identify potential pathways for therapeutic intervention. The adipose-derived hormone leptin acts via neurons in the brain that express the long form of the leptin receptor (LepRb) to decrease food intake and potentiate energy expenditure. Many aspects of leptin action via LepRb neurons of the medialbasal hypothalamus have been characterized, but they do not account for leptin's ability to modulate the incentive salience of food. A potential locus for this aspect of leptin action is the lateral hypothalamus (LHA), which contains Orexin (OX) neurons that project to and regulate the mesolimbic dopamine (DA) system to alter motivated behavior (including food intake and activity). My work in the laboratory of Dr. Martin Myers has begun to characterize LHA LepRb neurons, including a subpopulation that are discrete from, but synaptically connected with, OX neurons. This subpopulation of LHA LepRb neurons co-express opposing transmitters: the inhibitory neurotransmitter GABA and the excitatory neuropeptide neurotensin (Nts), a known regulator of the mesolimbic dopamine (DA) system;we therefore refer to this population as LHA LepRbNts neurons. I hypothesize that opposing GABA and Nts signaling from LHA LepRbNts neurons mediate distinct regulatory effects in energy homeostasis, particularly via the mesolimbic DA system. During the K99 mentored phase, I will explore this hypothesis in mice null for Nts signaling (Aim 1) while developing a novel mouse model (NtsFRT Neo-Cre mice) to selectively identify LHA LepRbNts neurons (Aim 2a) allowing for their functional interrogation. Dr. Myers has a substantial track record in creating novel mouse lines, and his mentorship during the K99 phase is essential for my ability to learn the molecular genetics skills necessary to create the NtsFRT Neo-Cre mice (and subsequent novel mouse models) that will be utilized for my independent research program. During the R00 phase, I will interbreed NtsFRT Neo-Cre mice and existing mouse models to generate mice that either selectively ablate all LHA LepRbNts neurons (Aim 2) or their GABA signaling (Aim 3) to identify the roles of these neurons overall and parse the specific role of GABA in energy homeostasis. Collectively, the scientific and career training facets of the mentored K99 phase will provide the tools to establish my independent research program in the R00 phase, supporting my transition to a productive research faculty position in the field of obesity. Overall, this line of research will determine the signaling mechanisms by which LHA LepRbNts neurons contribute to energy homeostasis, and their role in the pathogenesis of obesity PUBLIC HEALTH RELEVANCE: Obesity-linked diabetes is an increasing worldwide health problem, but limited understanding of the systems that regulate food intake and weight has hindered development of effective therapies for this disease. The studies herein will investigate the mechanisms by which a unique group of neurons in the lateral hypothalamus regulate food intake, weight, activity levels and hedonic or "craving" responses. This work will add to our understanding of the development of obesity, and characterization of these novel neurons may identify therapeutic intervention sites for treating obesity.